Guava MultiCaspase Assay
for detection of mid-stage apoptosis
The Guava® MultiCaspase Assay identifies cell populations in various stages of apoptosis by virtue of a fluorochrome-conjugated caspase inhibitor, sulforhodamine-valyl-alanyl-aspartyl-fluoromethylketone (SR-VAD-FMK).
Non-cytotoxic, cell permeant dye eliminates lysis and permeabilization procedures
Both cell permeable and non-cytotoxic, SR-VAD-FMK can readily cross cell membranes and covalently bind multiple caspase family members that have been activated during apoptosis. Unbound inhibitor is washed away, while covalently coupled reagent is retained inside the cell with the disabled caspases.
Simultaneous staining with the cell-impermeant dye, 7-AAD, serves to assess membrane structural integrity. 7-AAD is excluded from live, healthy cells and early apoptotic cells, but permeates later stage apoptotic and dead cells. The assay distinguishes four popoulations:
Live and early apoptotic (non-committed cells): SR-VAD-FMK (-) and 7-AAD (-)
Mid stage (committed) apoptotic cells: SR-VAD-FMK (+) and 7-AAD (-)
Late stage apoptotic and dying cells: SR-VAD-FMK (+) and 7-AAD (+)
Dead cells: SR-VAD-FMK (- or dim) and 7-AAD (+)
Key features
Tube and 96-well plate protocols
Works with both adherent and suspended cells
Analyzes apoptosis in single cells
FLICA reagents are non-cytotoxic and cell permeant, thus eliminating cell lysis and permeabilization procedures
Staining and data acquisition in ≤3 hours
Provides direct, absolute cell counts and population percentages
Data compatible with third party analysis software
Runs on Guava PCA, PCA-96, EasyCyte Mini, and EasyCyte Plus Systems
Ordering information
MultiCaspase Kit (PCA System) (50 tests)
4500-0101
MultiCaspase Kit (PCA-96, EasyCyte Mini and Plus Systems) (100 tests)
